Anti-Lactoferrin antibody [EPR30217-521R]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling Lactoferrin with ab325119 at 1/5000 dilution (0.01 ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were stained with anti-CD3 conjugated to Alexa Fluor®647. Then fixed with 4% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling Lactoferrin with ab325119 at 1/5000 dilution (0.01 ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were stained with anti-Gr-1 conjugated to Pacific Blue. Then fixed with 4% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling Lactoferrin with ab325119 at 1/5000 dilution (0.01 ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were stained with anti-CD11b conjugated to Brilliant Violet 421. Then fixed with 4% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Mouse bone marrow cells labelling Lactoferrin with ab325119 at 1/50000 (0.01 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in subset of CD11b+mouse bone marrows (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab8878 Anti-CD11b Rat monoclonal antibody was used to counterstain CD11b at 1/200 dilution, followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. (Magenta).

-ve control 1 : ab325119 at 1/50000 dilution, followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab8878 at 1/200 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Lactoferrin with ab325119 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on neutrophils in mouse spleen. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Immunohistochemical analysis of paraffin-embedded Mouse breast tissue labeling Lactoferrin with ab325119 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : brain

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Lactoferrin antibody [EPR30217-521R] (ab325119) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 77 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : liver

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Lactoferrin antibody [EPR30217-521R] (ab325119) at 1/1000 dilution

All lanes:

Mouse breast tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 77 kDa,36 kDa

false

Exposure time: 26s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactoferrin antibody [EPR30217-521R] (AB325119)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Lactoferrin with ab325119 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on mouse cerebrum. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins